Archiving a plasmid library on nitrocellulose

e4dna at att.net e4dna at att.net
Wed Dec 19 15:11:12 EST 2001


Hi! I am trying to archive a human cDNA library from 
overnight DH10B cultures without having to freeze each 
aliquot separately. I tried the Whatman CloneSaver Card 
system of spotting 5ul of the culture and then using a 
card punch to transform bacteria, but rarely recovered 
transformants. Either the plasmid DNA is being washed 
off or bacterial refuse on the card is inhibiting the 
transformations. Does anyone have suggestions on 
improving recovery with this system? 

Also, is it possible to completely release DNA from 
nitrocellulose or Nunc Nylon Membrane using salt elution 
instead of electroblotting? I want to try to use such 
filters for storing plasmids either from colonies that 
are grown directly on the filter or from cultures 
blotted onto them.

Thanks for your advice.
Adam  MGH  Boston

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